Journal of the American Society for Mass Spectrometry

, Volume 20, Issue 10, pp 1933–1943

Origin of supercharging in electrospray ionization of noncovalent complexes from aqueous solution


DOI: 10.1016/j.jasms.2009.06.012

Cite this article as:
Sterling, H.J. & Williams, E.R. J Am Soc Mass Spectrom (2009) 20: 1933. doi:10.1016/j.jasms.2009.06.012


The use of m-nitrobenzyl alcohol (m-NBA) to enhance charging of noncovalent complexes formed by electrospray ionization from aqueous solutions was investigated. Addition of up to 1% m-NBA can result in a significant increase in the average charging of complexes, ranging from ∼13% for the homo-heptamer of NtrC4-RC (317 kDa; maximum charge state increases from 42+ to 44+) to ∼49% for myoglobin (17.6 kDa; maximum charge state increases from 9+ to 16+). Charge state distributions of larger complexes obtained from heated solutions to which no m-NBA was added are remarkably similar to those containing small amounts of m-NBA. Dissociation of the complexes through identical channels both upon addition of higher concentrations of m-NBA and heating is observed. These results indicate that the enhanced charging upon addition of m-NBA to aqueous electrospray solutions is a result of droplet heating owing to the high boiling point of m-NBA, which results in a change in the higher-order structure and/or dissociation of the complexes. For monomeric proteins and small complexes, the enhancement of charging is lower for heated aqueous solutions than from solutions with m-NBA because rapid folding of proteins from heated solutions that do not contain m-NBA can occur after the electrospray droplet is formed and is evaporatively cooled.

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© American Society for Mass Spectrometry 2009

Authors and Affiliations

  1. 1.College of ChemistryUniversity of California-BerkeleyBerkeleyUSA